附子多糖对深低温保存腹主动脉线粒体结构和功能的影响  

Effects of Fuzi polysaccharide on structure and function of abdominal aorta mitochondria under cryopreservation

作　　者：周树兰 王成 ZHOU Shulan;WANG Cheng(Department of Hand Surgery,Fifth Affiliated Hospital of Zunyi(Zhuhai)Medical University,Zhuhai 519100,China)

机构地区：[1]遵义医科大学第五附属(珠海)医院手外科,珠海519100

出　　处：《山西医科大学学报》2022年第2期221-226,共6页Journal of Shanxi Medical University

基　　金：国家自然科学基金项目(81960824)。

摘　　要：目的探讨深低温保存中不同浓度附子多糖(Fuzi polysaccharide,FPS)对腹主动脉线粒体结构和功能的影响。方法显微镜下取48只SD大鼠腹主动脉,随机分为新鲜组、保存对照组(灭菌注射用水)、多糖保存组(FPS 0.1,1,10,20 mg/ml)。新鲜组取材后立即采用透射电镜观察线粒体内部超微结构变化,紫外-可见光分光光度计检测线粒体呼吸链复合体Ⅰ、Ⅱ、Ⅲ、Ⅳ活性,酶标仪检测ATP酶(Na^(+)-K^(+)-ATP酶、Ca ^(2+)-ATP酶)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-PX)、过氧化氢酶(CAT)活力和丙二醛(MDA)含量等指标对腹主动脉线粒体结构和功能进行综合评价;保存对照组和多糖保存组取腹主动脉后于-196℃液氮内保存1周后进行上述指标检测。结果与新鲜组比较,保存对照组线粒体结构完整性显著降低,复合体活性、ATP酶活力和抗氧化能力均大幅度下降(P<0.01),MDA含量明显升高(P<0.01)。与保存对照组比较,多糖保存组线粒体损伤减轻,复合体活性、ATP酶、抗氧化物酶活力增强和MDA含量降低,并从浓度1 mg/ml开始差异有统计学意义,当浓度达10 mg/ml时达峰值。结论深低温保存造成了线粒体结构和功能损害;附子多糖通过改善线粒体功能障碍减轻氧化应激损伤,具有保护腹主动脉线粒体结构和功能的作用。Objective To investigate the effects of different concentrations of Fuzi polysaccharide(FPS)on structure and function of abdominal aorta mitochondria during cryopreservation.Methods The abdominal aorta of 48 SD rats was taken under microscope,and randomly divided into fresh group,control group(sterilized water for injection)and polysaccharide preservation groups(FPS 0.1,1,10,20 mg/ml).The ultrastructural changes of mitochondria in fresh group were observed under transmission electron microscope(TEM)immediately after sampling,the activities of mitochondrial respiratory chain complexesⅠ,Ⅱ,ⅢandⅣwere detected by UV-visible spectrophotometer,and the activities of ATPase(Na^(+)-K^(+)-ATPase,Ca^(2+)-ATPase),superoxide dismutase(SOD),glutathione peroxidase(GSH-PX),catalase(CAT)and malondialdehyde(MDA)were detected by enzyme-labeled instrument.These indexes were used to evaluate the structure and function of abdominal aortic mitochondria.Abdominal aorta in control group and polysaccharide preservation group was taken and stored in liquid nitrogen at-196℃for one week,and then the above indexes were detected.Results Compared with fresh group,the integrity of mitochondrial structure,the activity of complexes,ATPase and the antioxidant capacity were significantly decreased in control group(P<0.01),while the content of MDA was increased significantly(P<0.01).Compared with control group,the damage of mitochondria was alleviated in polysaccharide preservation group,the activity of complexes,ATPase,and the antioxidant capacity were increased,and the MDA level was decreased,and the indexes peaked in 10 mg/ml group,and there was statistical difference between 1,10,20 mg/ml groups and control group.Conclusion Cryopreservation can cause the damage of mitochondrial structure and function.Fuzi polysaccharide can protect the structure and function of abdominal aorta mitochondria by improving the mitochondrial dysfunction and alleviating the oxidative stress injury.

关 键 词：附子多糖 深低温保存 腹主动脉 冷冻保护剂 线粒体损伤 

分 类 号：R318.5[医药卫生—生物医学工程]